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Histone H3 serine 57 and lysine 56 interplay in transcription elongation and recovery from S-phase stress.

Aslam A, Logie C - PLoS ONE (2010)

Bottom Line: Because phosphorylated human histone H3 serine 57 peptides have been detected by mass spectrometry we examined whether H3-S57 phosphorylation interplays with H3-K56 acetylation in vivo.Strikingly, opposite results were obtained in the context of a serine to alanine substitution at position 57 of histone H3.We speculate that histone H3-S57 couples H3-K56 acetylation to histone quaternary structures involving arginine 40 on histone H4 helix 1.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biology, Nijmegen Centre for Molecular Life Sciences, Radboud University Nijmegen, The Netherlands.

ABSTRACT

Background: Acetylation of lysine 56 of histone H3 plays an important role in the DNA damage response and it has been postulated to play an as yet undefined role in transcription, both in yeast and in higher eukaryotes. Because phosphorylated human histone H3 serine 57 peptides have been detected by mass spectrometry we examined whether H3-S57 phosphorylation interplays with H3-K56 acetylation in vivo.

Methodology/principal findings: To explore the physiological role of H3-S57, H3-K56 was mutated to mimic constitutively (un)acetylated forms of H3-K56 and these were combined with constitutively (un)phosphorylated mimics of H3-S57, in yeast. A phosphorylated serine mimic at position 57 lessened sensitivities to a DNA replication fork inhibitor and to a transcription elongation inhibitor that were caused by an acetylated lysine mimic at position 56, while the same substitution exacerbated sensitivities due to mimicking a constitutive non-acetylated lysine at position 56. Strikingly, opposite results were obtained in the context of a serine to alanine substitution at position 57 of histone H3.

Conclusions/significance: The phenotypes elicited and the context-dependent interplay of the H3-K56 and -S57 point mutations that mimic their respective modification states suggest that serine 57 phosphorylation promotes a nucleosomal transaction when lysine 56 is acetylated. We speculate that histone H3-S57 couples H3-K56 acetylation to histone quaternary structures involving arginine 40 on histone H4 helix 1.

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Interplay between H3-K56 and H3-S57 mutations.A. Five-fold serial dilutions of the indicated mutant strains were analyzed on the indicated YEPD plates. One representative experiment out of at least three is shown. Note the formamide sensitivity of both single H3-S57 mutants and also the inverse MMS, HU and 6-AU hypersensitivity relations between the H3-S57A and H3-S57E mutations in the contexts of the H3-K56R and H3-K56Q point mutations. B. Wild type H3, H3-S57A and H3-57E were tested as in (A) but using higher concentrations of MMS and of HU.
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pone-0010851-g003: Interplay between H3-K56 and H3-S57 mutations.A. Five-fold serial dilutions of the indicated mutant strains were analyzed on the indicated YEPD plates. One representative experiment out of at least three is shown. Note the formamide sensitivity of both single H3-S57 mutants and also the inverse MMS, HU and 6-AU hypersensitivity relations between the H3-S57A and H3-S57E mutations in the contexts of the H3-K56R and H3-K56Q point mutations. B. Wild type H3, H3-S57A and H3-57E were tested as in (A) but using higher concentrations of MMS and of HU.

Mentions: To uncover functional relations, we assessed phenotypes induced by compounds that target DNA damage to S-phase (MMS, HU), generate pyrimidine dimers (UV), disrupt hydrogen bonding (formamide) or that inhibit transcription (6-AU). Mutation of H3-S57 to alanine or glutamate had no effect on sensitivity to 6-AU, UV or growth at higher temperature (Figure 3A) and sensitized yeast to formamide to a similar extent. Interestingly however, the H3-S57E mutant was more sensitive than the H3-S57A mutant to DNA damage caused by MMS (Figure 3A). Additional experiments with higher doses of MMS and HU showed that both of the single H3-S57 substitutions were sensitive to S-phase stress. However, while H3-S57E was sensitive to both HU and MMS, S57A was not sensitive to HU, even at a higher concentration (Figure 3B). These results suggest that histone H3 serine 57 has some role when yeast cells face S-phase stress induced by MMS [34].


Histone H3 serine 57 and lysine 56 interplay in transcription elongation and recovery from S-phase stress.

Aslam A, Logie C - PLoS ONE (2010)

Interplay between H3-K56 and H3-S57 mutations.A. Five-fold serial dilutions of the indicated mutant strains were analyzed on the indicated YEPD plates. One representative experiment out of at least three is shown. Note the formamide sensitivity of both single H3-S57 mutants and also the inverse MMS, HU and 6-AU hypersensitivity relations between the H3-S57A and H3-S57E mutations in the contexts of the H3-K56R and H3-K56Q point mutations. B. Wild type H3, H3-S57A and H3-57E were tested as in (A) but using higher concentrations of MMS and of HU.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2877106&req=5

pone-0010851-g003: Interplay between H3-K56 and H3-S57 mutations.A. Five-fold serial dilutions of the indicated mutant strains were analyzed on the indicated YEPD plates. One representative experiment out of at least three is shown. Note the formamide sensitivity of both single H3-S57 mutants and also the inverse MMS, HU and 6-AU hypersensitivity relations between the H3-S57A and H3-S57E mutations in the contexts of the H3-K56R and H3-K56Q point mutations. B. Wild type H3, H3-S57A and H3-57E were tested as in (A) but using higher concentrations of MMS and of HU.
Mentions: To uncover functional relations, we assessed phenotypes induced by compounds that target DNA damage to S-phase (MMS, HU), generate pyrimidine dimers (UV), disrupt hydrogen bonding (formamide) or that inhibit transcription (6-AU). Mutation of H3-S57 to alanine or glutamate had no effect on sensitivity to 6-AU, UV or growth at higher temperature (Figure 3A) and sensitized yeast to formamide to a similar extent. Interestingly however, the H3-S57E mutant was more sensitive than the H3-S57A mutant to DNA damage caused by MMS (Figure 3A). Additional experiments with higher doses of MMS and HU showed that both of the single H3-S57 substitutions were sensitive to S-phase stress. However, while H3-S57E was sensitive to both HU and MMS, S57A was not sensitive to HU, even at a higher concentration (Figure 3B). These results suggest that histone H3 serine 57 has some role when yeast cells face S-phase stress induced by MMS [34].

Bottom Line: Because phosphorylated human histone H3 serine 57 peptides have been detected by mass spectrometry we examined whether H3-S57 phosphorylation interplays with H3-K56 acetylation in vivo.Strikingly, opposite results were obtained in the context of a serine to alanine substitution at position 57 of histone H3.We speculate that histone H3-S57 couples H3-K56 acetylation to histone quaternary structures involving arginine 40 on histone H4 helix 1.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biology, Nijmegen Centre for Molecular Life Sciences, Radboud University Nijmegen, The Netherlands.

ABSTRACT

Background: Acetylation of lysine 56 of histone H3 plays an important role in the DNA damage response and it has been postulated to play an as yet undefined role in transcription, both in yeast and in higher eukaryotes. Because phosphorylated human histone H3 serine 57 peptides have been detected by mass spectrometry we examined whether H3-S57 phosphorylation interplays with H3-K56 acetylation in vivo.

Methodology/principal findings: To explore the physiological role of H3-S57, H3-K56 was mutated to mimic constitutively (un)acetylated forms of H3-K56 and these were combined with constitutively (un)phosphorylated mimics of H3-S57, in yeast. A phosphorylated serine mimic at position 57 lessened sensitivities to a DNA replication fork inhibitor and to a transcription elongation inhibitor that were caused by an acetylated lysine mimic at position 56, while the same substitution exacerbated sensitivities due to mimicking a constitutive non-acetylated lysine at position 56. Strikingly, opposite results were obtained in the context of a serine to alanine substitution at position 57 of histone H3.

Conclusions/significance: The phenotypes elicited and the context-dependent interplay of the H3-K56 and -S57 point mutations that mimic their respective modification states suggest that serine 57 phosphorylation promotes a nucleosomal transaction when lysine 56 is acetylated. We speculate that histone H3-S57 couples H3-K56 acetylation to histone quaternary structures involving arginine 40 on histone H4 helix 1.

Show MeSH
Related in: MedlinePlus